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Evaluation Studies
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Quantitative analysis of Howell-Jolly bodies in children with sickle cell disease.
Experimental Hematology 2007 Februrary
OBJECTIVES: Although functional asplenia in sickle cell disease (SCD) begins early in life and has important clinical consequences, quantitative measurement of splenic function is not readily available. A novel high-throughput flow cytometric method for quantitating Howell-Jolly bodies (HJB) has been developed which isolates HJB-containing CD71(+) and CD71(-) erythrocytes. Analysis of these cell populations allows quantitative measurement of splenic filtrative function and possible chromosomal damage.
METHODS: Blood specimens from 147 children with SCD were analyzed using a high-throughput flow cytometric method. Enumeration of the following populations was accomplished: 1) CD71(+) reticulocytes among total erythrocytes, identifying the youngest erythroid cell population; 2) HJB-containing CD71(+) reticulocytes, which isolate young erythrocytes containing micronuclei as an index of cytogenetic damage; and 3) HJB-containing CD71(-) erythrocytes, identifying older erythrocytes containing micronuclei, indirectly measuring splenic function.
RESULTS: Children with HbSC (n = 24) had slightly elevated HJB frequencies, while children with HbSS (n = 125) had highly elevated frequencies within CD71(+) cells (0.44% +/- 0.40%, normal 0.12% +/- 0.06%, p < 0.001) and CD71(-) cells (2493 +/- 2303 per million RBC, normal 20 +/- 11, p < 0.001). Using a multiple regression model, the frequency of HbSS CD71(+) reticulocytes containing HJB was significantly influenced by hydroxyurea use (p < 0.0001), age (p = 0.0288), and splenectomy (p = 0.0498). Similarly, mature CD71(-) erythrocytes containing HJB were positively correlated with hydroxyurea (p = 0.0001), age (p < 0.0001), and splenectomy (p = 0.0104).
CONCLUSIONS: HJB quantitation by flow cytometry is a novel assay for measuring splenic function and may be valuable for investigating the efficacy and safety of therapeutic options for children with SCD.
METHODS: Blood specimens from 147 children with SCD were analyzed using a high-throughput flow cytometric method. Enumeration of the following populations was accomplished: 1) CD71(+) reticulocytes among total erythrocytes, identifying the youngest erythroid cell population; 2) HJB-containing CD71(+) reticulocytes, which isolate young erythrocytes containing micronuclei as an index of cytogenetic damage; and 3) HJB-containing CD71(-) erythrocytes, identifying older erythrocytes containing micronuclei, indirectly measuring splenic function.
RESULTS: Children with HbSC (n = 24) had slightly elevated HJB frequencies, while children with HbSS (n = 125) had highly elevated frequencies within CD71(+) cells (0.44% +/- 0.40%, normal 0.12% +/- 0.06%, p < 0.001) and CD71(-) cells (2493 +/- 2303 per million RBC, normal 20 +/- 11, p < 0.001). Using a multiple regression model, the frequency of HbSS CD71(+) reticulocytes containing HJB was significantly influenced by hydroxyurea use (p < 0.0001), age (p = 0.0288), and splenectomy (p = 0.0498). Similarly, mature CD71(-) erythrocytes containing HJB were positively correlated with hydroxyurea (p = 0.0001), age (p < 0.0001), and splenectomy (p = 0.0104).
CONCLUSIONS: HJB quantitation by flow cytometry is a novel assay for measuring splenic function and may be valuable for investigating the efficacy and safety of therapeutic options for children with SCD.
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